1. Technical Field
The present disclosure relates to a container for performing nucleic acid amplification reaction therein. More particularly, the present disclosure relates to a reaction tube for performing isothermal polymerase chain reaction therein.
2. Description of Related Art
The nucleic acid amplification reaction is a scientific technique in molecular biology to amplify a single or a few copies of a particular deoxyribonucleic acid (DNA) sequence by repeating the same procedure with particular polymerases. The common techniques such as polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), and real-time polymerase chain reaction (real-time PCR) all belong to nucleic acid amplification reaction techniques.
The PCR is majorly used to amplify a particular DNA, whereas the RT-PCR is used to reverse transcribing a specified RNA fragment to a particular DNA fragment followed by amplifying the particular DNA fragment, namely complementary DNA (cDNA). The real-time PCR, also called quantitative PCR, is used to amplify and quantify a targeted DNA simultaneously, where the main reagents associated in this procedure are fluorescent probe and dyes. Taking all together, the principle of the nucleic acid amplification reactions mentioned above is PCR.
Furthermore, some skills presented lately also belong to nucleic acid amplification reactions, such as rolling circle amplification (RCA), loop mediated amplification (LAMP), nucleic acid sequence based amplification (NASBA), and three way junction (TWJ).
Regarding to general PCR, the initialization step is used for mixing and heating DNA templates, primers, and a buffer solution to the reaction temperature about 90° C. for disrupting the hydrogen bonds between two single-stranded DNA templates, namely the denaturation step. The second step is used for cooling the reaction temperature to about 50° C. for annealing the primers and the single-stranded DNA template. The final step is used for holding the temperature at about 70° C. for extending the primers. The particular DNA is copied by repeating the above procedure.
The types of the apparatus for the nucleic acid amplification reaction are classified according to the prices. The cheaper type includes a container, such as a tube or a capillary, and two heaters. The two heaters are respectively disposed on the two ends of the container. One heater heats the container to about 90° C., and the other heats the container to about 50° C. The solution convection in the container takes place because of the density difference of the solution at the two ends of the container, wherein the density difference is caused by the temperature difference between the two ends. The DNA and the primers is circulated through the container and heated from 90° C. to 50° C. circularly for performing the nucleic acid amplification reaction.
The heater is made of a metal block. The block has a groove for receiving the end of the container, and the shape of the groove is designed to be fitted to the end of the container. However, the groove does not completely match with the container; it implies that the groove remains some protrusions and indentations when the end of the container is received by the groove. Therefore, the edge of the protrusions and the indentations do not completely and evenly contact to the container in order to conduct heat to the container. Thus, the container cannot be heated evenly. The reaction efficiency of the nucleic acid amplification reaction will be reduced.